SU652901A3 - Biomass obtaining method - Google Patents

Description

This invention relates to a bacteriological cell mass production technique containing nrotein, as well as fats, carbohydrates and vitamins, and is used as a basic substance for feed and food products. A known method for producing biomass involves growing bacteria of the genus Methvlomonas under conditions of aeration on nitrous medium containing methanol as a carbon source, nitrogen source and mineral salts 1. This method is the closest to the proposed technical essence and the achieved result. The disadvantage of this method is not high enough biomass yield. The aim of the invention is to increase the biomass yield. For this, according to the proposed method of the genus Methylomonas, Methylomonas clara ATCC 31226 is used, while the concentration of methanol in the medium is maintained between 0.5–15 mg%, preferably 0.5–3 mg%. The method is described as follows. Bacteria 1 of tamma L e1Bu1topo5 clara ATCC 31226 are grown under aerobic conditions in a nutrient medium containing methanol at a concentration of 5-100 ppm, preferably 5-30 ppm, of nitrogen, such as potassium nitrate, ammonium sulfate, ammonium phosphate and ammonia, minerals, such as di1n; potassium phosphate or disodium phosphate, as well as magnesium and potassium salts, iron salts, copper, molybdenum, contained in the water-conducting water in the form of trace elements. The process is carried out periodically or continuously at temperatures between 30 and 42 ° C (mainly between 35-39 ° C, in agitated fermenters, and also in loop reactors. The liquid pest medium in the fermenter is blown with air in an amount of 0.1-1.5 liter of air per 1 liter of nutrient medium per 1 minute (in.m). The concentration of methanol is maintained continuously, for example, by measuring the flow rate of nitrogen, the proportion of biomass in suspension, or.

Preferred by measuring carbon dioxide emissions. Thanks to this, the exactness of the dosage of i1aYuT is the dosing of iodaci meta1 ol with a quick reaction to the irradiation of the residue of g.n with a few R 1.1 de, 1 e and e e glo to c and i from. V i) and BJ e concentration of methanol is determined by measuring the amount of gaseous-1-o methanol and OOMO1C1 and. amine ionization detector.

The pH value of the culture culture, the state of titatie. Green and growing biomass, is maintained between 4.0 and 9.0, usually between 6.0 and 7.2, by adding the appropriate alkali, such as caustic soda or ya.li, and; n1 acid, nanopolymer hydrochloric or sulfuric.

Biomass extraction is carried out in a manner known per se, e.g. centrifuging () by bending it with water), water, and a paste-like bomase containing 75% OO / O water is obtained.

Drying of the base is carried out with the help of, the most | imer, roller sui1.1.1ki, dryers with a cistern bed or a rotary dryer.

The product contains 1 to 4 wt. ° / c1 water and 80-90 ve. (o) crude nrotei: ca, in this, in a crude nano-dark, the proportion of amino acids is 75-80% by weight, and

C1) edible amino acids about 50 weight. %

of the total amino acid content.

The content of nucleic acids is 10–14 wt./o, and the fat is 510 wt. /about

and carbohydrates - 5 - K) weight. ° / o.

Biomass (dry) isg contains pigments, tokiechechny veychchv, as well as slnzi and nolioks1 mas.l nig of acid, smelling substances and second-) metabolites.

A new species, Mcthylonionas clara ATCC 31226, is described as follows.

11o1 adenne growth. Does not dodge on | -,: || okom agar or glucose ereda, agar, sleep, or nitatel medium on m boulevon, as well as platinum, nentone agar or non-1 ton nitates. , litmus milk. Digs i-: a siitsticheskoy iitate, liyu medium containing .methanol.

Morpho.logn. Short paws 0.5-1 .5 / C, motile due to the nostril rsch1x ciliary any formation of snor, cysts.

The shape of the colonies is round, transparent, with light gloss. Pigments do not form.

Physiologists. Digs at 20–25 ° C, 30–35 ° C, 37–39 ° C and 41 ° C. Optimum temperature, optimal pH 6.8 - 7.0, gram-negative.

Growth factors. Does not form nanoxy butyric acids, indole, acetone,. does not liquefy gelatins, does not hydrolyze starch, does not form lemonic acid, does not coagulate milk, does not grow on ureas, acetate, trimethylamine, monomethylamine, formate, formaldehyde, sugar, polysaccharides, alcohols (except methanol).

NOx, cytochrome oxide azu, catalase, isocitrate dehydrogenase, malate dehydrogenase. oxidase, grows on ammonium salts, dimethylamine, fixes Xs, CO ,.

fipUMip 1. Schlamm aletfiyloiiioHa. cbiia FII B 5460 ATCC 31226, flared 111, ayut tubules c.:leduyülceiCHJ c;) c18 g; From Ra (850 / o) 2,0 ml; Nl-UOM (120 / o-ny) 3.0 ml:

 0.01r; H2SCi l, 2r: MgSf X X7ND) - 0.8 g; FeSO, 7HiO 0.03 g, methanol (additive in front of the tubes) - 10.0 ml: a trace element solution containing CaCO3. NZVOZ, .MnSO4, HNZSL, Na.AlaCV - 1.0 ml: vada - I l: p1 ie) with sterilization 6,7.

The slopes of the 1st tube with this compound ia are heated for 30 minutes in an autoclave with, and then they infect MethYloHioiias clara and incubate for two days and 37 ° C. Of the two tubes 10 m; | fnzio, chogical solution of cooked so.ln B1) 1 wash the cell mass and inoculate on the next CTy ieiiH. This stage - granular Ku. Eltu1) a (forculture). Use 2 liters of Er bulb. Lenmeyer, 1 l of the specified nutrient solution, but without arajia. To the solution are added 3.3 ml sterile filtered; methanol. For 3 days in: 15 ° 37 ° C (with vlabl) H anime npii 220 rpm and amnlite de 4 cm.

After 24 hours and 48 hours, add 3.3 ml m with La NOL a.

The trace, y1On., Th stage (basic l.Lu1) a) is carried out in a 25 l fermenter filled with 20 l of the indicated nilate; || HHH) solution |) a. but without methanol and agar.

After sterilization in (jiepMeirrep, 2, h (| k) handles and (fermentation nro1)) are given at the table and bladed mei1a, lk iri 500 rpm, 37 C, air rest) 0 l / min 0, 5 vvn, pressure 0.02, pH 6.6.

Melanool. L. Heading 20 m, ln each time with 1 adeium) 1 B1) | division of coal, very few lots add 20. Ml of methanol, and more than 1% 0.1 vol. / ;. -1 through 22 h 20 l of the suspension main ku,) s no. Pouring; perspiration in the farm 1 gge) 200 l

c, k) i and (|) srm (41 tatsi1: those 1 of the 37Cj npo; iyp, Ka WHO'LUSHER 6 8 m-; h, 0.5 - 0.75 VVM: pressure 0.02 Mia: speed deformation-i15aii - 380 (; b / mip: p11 supportive; 1n () th Yu / o-amy miah water 6.7 6.8.

The flow of melio.la is determined by measuring its co-profile in the air using and, la:; a special-ionization detector. moreover, n) and after the amount of melano.la is below 50 him, s. follows. the addition of methanol.

With 600 him of methanol in the exhaust air in the ellasor, it contains 0.22 / o methane, l.

After 20 ij fermentation, 200 l of a forfermeiter suspension is added to the main fermenter, which contains 2 thousand .i nutrient solution and has the following operating conditions: temperature 37 ° C; air blow 60-80 0.5-0.75 inrm; pressure - 0.02 MPa; speed of change; 1 170 rpm; pH - 6.7.

The methanol feed was carried out in the same manner as in a 200 L formenter. The copolymer of methanol in the nutrient solution is im.

After 22 hours of fermentation, the cell mass is separated, for which the pH is adjusted to 4.0 by the addition of dilute sulfuric acid. Cell mass is separated on separators at 400 rpm. Separation can also be carried out at pH 6.5-6.8.

The separated cell mass with a dry matter content is washed with water and the separator is adjusted to a dry content of 25 ° / o.

After that, the cell mass is dried in a spray dryer at an inlet temperature of 120-150 ° C. Received ioroyuk contains, weight. ° / o: 1.5-3.5 residual moisture Mi (N 6.25) crude protein 85; Aminokilot 74; Edible amino acids 50, nucleic acids - 8--12; raw fat - 6-8, raw ash - 5-6.

Example 2. Strain Alethylomonas elara FH-B-5460 ATSS 31226 grown as in example 1.

A 3000 liter main fermenter is continuously air blown. Fermentation conditions;

Temperature, ° С37

Air blowing, 80.1 0.67 inrm

Pressure, MPa0,01

Speed, rpm 390

The pH value is 6.8.

200 l of the culture suspension from the fermenter are poured into the main fermenter containing 2000 l of nutrient solution (as in example 1, but without agar). The concentration of methanol is measured and adjusted by adding a mixture of methanol and water in a volume cooTHOHieniiii 40:60, so that the average concentration of methano.ch is maintained in a jiacTBope feeder of 10 p 50 ipm.

After preparation, 7–10 kg; l for the meter, 1 1 hour continuous operation).

The nutrient medium is added. 1 at a rate of 0.25 D for hours, about 12 hours later, the atoms are increased to D 6.33 hours, which is about 650 liters / hour. After this time period is established, the flow balance is established. The average concentration of free methanol is now maintained between 10 and 20 him. The appropriate amount of nutrient medium with grown to, 1-unit weight Was withdrawn for 12 hours, kept in an intermediate tank without the addition of methanol, and then Vedht cell separation and suprule as in example 1.

The cell mass thus obtained contains 2-4% of residual moisture and

0 crude protein (N 6.25) 81 / o, amino | Isl (:) t, C11 edible amino acids 50 / o of the total content of amino acids, nucleic acids 8-, crude fat 5-10 ° / o, crude ash 5-.

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Form of Invention

The method of obtaining bio: mass, ired) watching the BiiipauuiBaHHc bacterium 11 of Methylomonas in the form of aeration on the substrate, containing methanol as a carbon source, a source of nitrogen and minerals1) Salt, which is excellent, because with the aim of newer release, , ii3 X) and Methyloiiionas use H1Tamm .V ethylomonas clara LTCC 31226. while the end of methanol in the medium is maintained at 0.5–15 mg, and preferably 0.5–3.

Petochiii information taken into consideration at the examination

1. Patent of Great Britain L 1420-145, cl. C 12 D 13:06, 07.01.76.